ArticleInternational journal for parasitology2012
Flow cytometry-assisted rapid isolation of recombinant Plasmodium berghei parasites exemplified by functional analysis of aquaglyceroporin.
Article in International journal for parasitology, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed.
- Eliminating malaria transmission requires targeting immature and mature gametocytes through lipoidal uptake of antimalarials.Nature communications · 2024Article
- Novel Ion Channel Genes in Malaria Parasites.Genes · 2024Review
- IntracellularProceedings of the National Academy of Sciences of the United States of America · 2023Article
- Methodology to streamline flow cytometric-based detection of early stage Plasmodium parasitemia in mice.Journal of microbiological methods · 2022Article
- Absence of PEXEL-Dependent Protein Export inFrontiers in genetics · 2021Article
- An apicoplast-resident folate transporter is essential for sporogony of malaria parasites.Cellular microbiology · 2021Article
- Overlapping and distinct roles of CDPK family members in the pre-erythrocytic stages of the rodent malaria parasite, Plasmodium berghei.PLoS pathogens · 2020Article
- Application of the Brown dynamics fluctuation-dissipation theorem to the study of Plasmodium berghei transporter protein PbAQP.Frontiers in physics · 2020Article
- An in silico down-scaling approach uncovers novel constituents of the Plasmodium-containing vacuole.Scientific reports · 2018Article
- Aquaglyceroporin PbAQP is required for efficient progression through the liver stage of Plasmodium infection.Scientific reports · 2018Article
- Rapid inducible protein displacement inWellcome open research · 2017Article
- Distinct Prominent Roles for Enzymes of Plasmodium berghei Heme Biosynthesis in Sporozoite and Liver Stage Maturation.Infection and immunity · 2016Article
- Phylogenetic profiles of all membrane transport proteins of the malaria parasite highlight new drug targetsMicrobial cell (Graz, Austria) · 2016Article
- Efficient monitoring of the blood-stage infection in a malaria rodent model by the rotating-crystal magneto-optical method.Scientific reports · 2016Article
- Functional profiles of orphan membrane transporters in the life cycle of the malaria parasite.Nature communications · 2016Article
- The Plasmodium berghei translocon of exported proteins reveals spatiotemporal dynamics of tubular extensions.Scientific reports · 2015Article
- Conditional Degradation of Plasmodium Calcineurin Reveals Functions in Parasite Colonization of both Host and Vector.Cell host & microbe · 2015Article
- Towards genome-wide experimental genetics in the in vivo malaria model parasite Plasmodium berghei.Pathogens and global health · 2015Review
- A rapid and robust selection procedure for generating drug-selectable marker-free recombinant malaria parasites.Scientific reports · 2014Article
- Identification of vital and dispensable sulfur utilization factors in the Plasmodium apicoplast.PloS one · 2014Article
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4 authors.
Funding
Abstract
The most critical bottleneck in the generation of recombinant Plasmodium berghei parasites is the mandatory in vivo cloning step following successful genetic manipulation. This study describes a new technique for rapid selection of recombinant P. berghei parasites. The method is based on flow cytometry to isolate isogenic parasite lines and represents a major advance for the field, in that it will speed the generation of recombinant parasites as well as cut down on animal use significantly. High expression of GFP during blood infection, a prerequisite for robust separation of transgenic lines by flow cytometry, was achieved. Isogenic recombinant parasite populations were isolated even in the presence of a 100-fold excess of wild-type (WT) parasites. Aquaglyceroporin (AQP) loss-of-function mutants and parasites expressing a tagged AQP were generated to validate this approach. aqp(-) parasites grow normally within the WT phenotypic range during blood infection of NMRI mice. Similarly, colonization of the insect vector and establishment of an infection after mosquito transmission were unaffected, indicating that AQP is dispensable for life cycle progression in vivo under physiological conditions, refuting its use as a suitable drug target. Tagged AQP localized to perinuclear structures and not the parasite plasma membrane. We suggest that flow-cytometric isolation of isogenic parasites overcomes the major roadblock towards a genome-scale repository of mutant and transgenic malaria parasite lines.
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